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Germ cell-specific decrease of acrosomal proteolytic activity, sperm motility, and number in mitomycin C-treated mice.

Assessment of mammalian sperm acrosomal proteolytic activity, sperm motility, and sperm count may be useful for detecting mutagens, carcinogens, developmentally active agents, and antifertility effects. Groups of six albino mice were given a single i.p. injection of 5 mg/kg mitomycin C (MC) or saline. One treated and one control group of mice were killed 1, 3, 5, 7, or 10 weeks later. Sperm extracted from the vasa deferentia at these killing times were derived from cells treated as spermatozoa, spermatids, preleptotene-late spermatogonial cells, spermatogonial cells, and spermatogonial stem cells. In sperm derived from treated preleptotene or spermatogonial cells, the sperm count, sperm motility, and acrosomal proteolytic activity were decreased significantly. Acrosomal proteolytic activity was also decreased in sperm from spermatogonial stem cells. None of these sperm phenotypes were decreased in treated spermatozoa and spermatids. We propose the hypothesis that induced loss of sperm motility and acrosomal proteolytic activity in single spermatozoa derived from MC-treated spermatogonial cells is caused by mutational or developmental effects, whereas in preleptotene-derived and late-spermatogonium-derived sperm similar dysfunction results from developmental effects. Our data support the hypothesis indirectly. Since a low sperm count is correlated with decreased fertility and acrosomal proteolytic activity is essential for penetration of the zona pellucida by the sperm, the presence of these sperm phenotypes may help to detect chemicals with antifertility effects.

Acrosin↗

Reference value of semen quality in Chinese young men.

The purpose of the study was to evaluate semen quality in young Chinese men and to establish reference values. Normal healthy young men from seven geographical areas were enrolled. The study showed that the mean sperm volume was 2.61 mL, and mean percent of sperm with forward progression was 59.89, while median of semen viability was 79.0%, and geometric mean of semen density was 55.45 x 10(6)/mL. Proportion of routine semen indexes that met World Health Organization (WHO) criteria were as follows: 81.9% for semen volume, 91.1% for liquefaction time, 93.4% for viscosity, 90.8% for pH, 81.3% for sperm with forward progression, 65.3% for sperm viability, 93.8% for semen density, 98.8% for normal sperm morphology, and 89.1% for total sperm count. Participants whose sperm met all WHO standard parameters accounted for 42.3%. Because the infertility rate in China is about 10-15%, the fifteenth percentile of semen parameters might be used as the lower limit of reference values, which may be more appropriate for young Chinese men. The fifteenth percentiles of parameters in this study were as follows: 1.5 mL for semen volume, 7.2 for pH value, 45% for proportion of sperm with forward progression, 68% for sperm viability, 30 x 10(6)/mL for semen density, 68% for proportion of sperm with normal morphology, and 50 x 10(6) for total sperm count.

Adult↗

Semen quality of spinal cord injured men is better when obtained by vibratory stimulation versus electroejaculation.

PURPOSE: Most spinal cord injured men require assisted ejaculation procedures to obtain semen, and the majority can achieve this result by vibratory stimulation or electroejaculation. We determined if semen obtained by vibratory stimulation differed in quality from that obtained by electroejaculation. MATERIALS AND METHODS: Between subjects and within subjects designs were used. Of 77 spinal cord injured men 23 underwent vibratory stimulation only, 44 electroejaculation only and 10 both procedures. Antegrade, retrograde and total ejaculates were analyzed in each subject for total sperm count, percent motile sperm and percent sperm with rapid linear motion. RESULTS: With vibratory stimulation compared to electroejaculation the percent motile sperm and percent sperm with rapid linear motion were significantly greater, whereas total sperm count was similar, in the antegrade specimens and total ejaculates. This finding was true for different groups of subjects as well as within a group of the same subjects. CONCLUSIONS: Semen obtained by vibratory stimulation is of better quality than that obtained by electroejaculation. In medical practices that include assisted ejaculation of spinal cord injured men, we recommend obtaining a specimen by vibratory stimulation. If that method fails electroejaculation should be performed.

Adult↗

The impact of calcium, magnesium, zinc, and copper in blood and seminal plasma on semen parameters in men.

To investigate the impact of calcium, magnesium, zinc, and copper in blood and seminal plasma on semen parameters, 107 fertile and 103 subfertile males provided a standardized blood and semen specimen. Total calcium and magnesium concentrations were determined with colorimetric end point assay procedures. Zinc and copper were determined by flame atomic absorption spectrophotometer (AAS). Semen analysis was performed according to World Health Organization guidelines (1992). The concentrations of calcium, magnesium, zinc, and copper in blood and seminal plasma were not different between the subfertile and fertile group. Weak correlations were demonstrated between blood plasma zinc concentrations and sperm count (rs = 0.18), sperm motility (rs = 0.15), and abnormal sperm morphology (rs = 0.13). Zinc and magnesium concentrations in seminal plasma correlated weakly with sperm count (rs = 0.17 and rs = 0.16, respectively), and copper concentrations in blood plasma with motility (rs = 0.25). Strong correlations were found between calcium, magnesium, and zinc in seminal plasma. Although calcium, magnesium, zinc, and copper play an essential role in spermatogenesis and fertility, the determination of these elements in blood and seminal plasma does not discriminate on the basis of fertility in this group of men.

Adult↗

Decreased sperm number and motile activity on the F1 offspring maternally exposed to butyl p-hydroxybenzoic acid (butyl paraben).

Butyl p-hydroxybenzoic acid (butyl paraben, BP) is widely used as a preservative in food and cosmetic products. Routledge et al showed that BP is weakly estrogenic in both in vitro and in vivo (rat uterotrophic) analyses. We investigated whether maternal exposures to BP during gestation and lactation periods affected the development of the reproductive organs of the F1 offspring. Pregnant Sprague-Dawley rats were injected subcutaneously with 100 or 200 mg/kg of BP from gestation day (GD) 6 to postnatal day (PND) 20. In the group exposed to 200 mg/kg of BP, the proportion of pups born alive and the proportion of pups surviving to weaning were decreased. The body weights of female offspring were significantly decreased at PND 49. The weights of testes, seminal vesicles and prostate glands were significantly decreased in rats exposed to 100 mg/kg of BP on PND 49. In contrast, the weights of female reproductive organs were not affected by BP. The sperm count and the sperm motile activity in the epididymis were significantly decreased at doses of 100 and 200 mg/kg of BP. In accordance with the sperm count in the epididymis, the number of round spermatids and elongated spermatids in the seminiferous tubule (stage VII) were significantly decreased by BP. Testicular expression of estrogen receptor (ER)-alpha and ER-beta mRNA was significantly increased in 200 mg/kg of BP treated group at PND 90. Taken together, these results indicated that maternal exposure of BP might have adverse effects on the F1 male offspring.

Animals↗

Effects of purification or split ejaculation of semen and stimulation of spermatozoa by caffeine on their motility and fertilizing ability with the use of zona-free hamster ova.

Purification of semen with Sephadex G-50 medium powder or bovine serum albumin (BSA) (20% in medium) columns increases the percentage of motility. Spermatozoa that passed a BSA column or a Sephadex column have significantly higher velocity than unmanipulated spermatozoa. Caffeine (7 mM) stimulation did not show any effect on motility. Neither purification procedures nor stimulation with caffeine improved the in vitro fertilization capacity of capacitated spermatozoa, as determined by interaction with zona-free hamster ova. No difference in in vitro fertilizing capacity could be demonstrated between the first and the last fractions of split ejaculates. After grouping the fractions according to sperm counts, fractions with higher total (motile) sperm counts, and fractions with higher sperm counts had significantly higher fertilization percentages than the fractions with lower (motile) sperm counts.

Animals↗

Repeated analysis of semen parameters in beagle dogs during a 2-year study with the HMG-CoA reductase inhibitor, atorvastatin.

Sperm analyses are often incorporated into reproductive toxicity studies in rats. Due to the relative ease of collecting multiple samples throughout a study, semen analysis in non-rodents such as dogs offers the opportunity to assess potential development of functional effects of compounds on male reproduction over time. In the present study, semen parameters were evaluated in beagle dogs during and at termination of a chronic toxicity study with the 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor, atorvastatin. Male dogs received 0, 10, 40, or 120 mg/kg orally in gelatin capsules for up to 104 weeks (n = 10/group). After 52 weeks of dosing, 3 dogs/group were euthanized, and 2/group were withdrawn from treatment for a 12-week reversal period and euthanized at Week 64. The remaining 5/group continued treatment until Week 104. Semen was collected from all animals for 3 consecutive weeks prior to termination of the 52-week animals (Weeks 50, 51, 52) for analysis of sperm parameters, using manual methods of evaluation. Semen was collected from the remaining animals at Weeks 64, 78, 91, and 104, and was analyzed. At necropsy, testes, epididymides, and prostates were weighed and evaluated histologically, and epididymal sperm counts were determined. Serum cholesterol was decreased 25--60% at all doses during the study. There were no drug-related differences in semen volume and color, total sperm count, and sperm concentration, morphology, progressiveness, and percent motility during treatment with atorvastatin. There were also no effects on reproductive organ weights or histopathology, and no effects on epididymal sperm count. Thus, incorporation of semen analyses into this study allowed the evaluation of potential male reproductive effects in dogs at multiple time points during the study. Statistical power calculations demonstrated acceptable statistical power (> 80%) for semen sperm count, concentration, morphology, and motility with group sizes of 8--10 animals, and for semen sperm count and concentration or epididymal sperm count with group sizes of 3--5 animals, using the methodology described in this paper.

Acyl Coenzyme A↗

Effects of in utero exposure to DES on male progeny.

Diethylstilbestrol (DES) was prescribed for almost 30 years by physicians who believed the nonsteroidal synthetic estrogen was beneficial in both the treatment of threatened abortions and the prevention of spontaneous abortions. Toxemia, premature delivery, postmaturity, and stillbirth were said to be significantly reduced in patients who were treated with DES. The consequences of in utero exposure to DES on female progeny have been widely publicized. However, male progeny also suffer the effects of in utero exposure to DES. These effects include epididymal cysts, small penile size, and cryptorchidism. Altered reproductive capacity is suggested by diminished sperm counts and sperm penetration assays.

Abnormalities, Drug-Induced↗

Treatment of men with oligoasthenozoospermia and asthenozoospermia with kallikrein.

A randomised group of 75 men with idiopathic oligoasthenozoospermia and asthenozoospermia were subjected to control study with kallikrein. Kallikrein was given to 45 men 600 units daily in a period of 3-6 months. The other group of 30 infertile men was followed during the same period of time. Percentage of motile sperm was significantly improved after 3-6 months treatment with kallikrein. Sperm count and percentage of morphologically normal sperm showed improvement but was not statistically significant. Infertile men with sperm count below 20 X 10(6)/ml had greater improvement in sperm count than men with sperm count over 20 X 10(6)/ml. This study emphasizes that kallikrein is a relatively good drug for restoring sperm motility in idiopathic oligoasthenozoospermia and asthenozoospermia.

Adult↗

Application of a bovine cervical mucus penetration test.

One hundred seventy-eight ejaculates obtained from infertility patients were evaluated by routine semen analysis and by a bovine cervical mucus penetration test (BCMPT). A significant correlation (p less than 0.01) was observed between mucus penetration and both sperm count (r = 0.349) and sperm motility (r = 0.394). One hundred fifty-two of 178 patients (85%) had normal sperm counts (greater than 20 x 10(6)/ml). Of these patients, 68% had good (greater than 30 mm), 26% had questionable (21-30 mm), and 7% had abnormal (less than 20 mm) penetration values. One hundred sixty-one of 178 patients (90%) had normal sperm motilities (greater than 40%). Of these patients, 71% had good, 25% had questionable, and 4% had abnormal penetration values. Conversely, 46% and 18% of patients with abnormal sperm count and motility, respectively, had normal penetration values (greater than 30 mm). A significant relationship (p less than 0.05) was observed between the BCMPT and pregnancy problems not apparent by semen analysis data, and may prove to be a useful adjunct to the use of routine semen analysis in evaluating male fertility.

Animals↗

Varicocele and the morphology of spermatozoa.

It is believed that varicocele is characteristically associated with specific abnormalities of spermatozoal morphology (increased numbers of tapered and amorphous forms). This report summarizes the results of a prospective study of spermatozoal morphology in 208 male partners of infertile couples seen consecutively during an 18-month period. After exclusion of 23 azoospermic males, sperm morphology in the remaining 56 patients with varicocele and 129 men without varicocele were compared. Although the incidence of abnormal spermatozoa was higher in patients with varicocele, no significant differences in the patterns of morphology were observed. In both groups of patients the percentage of normal spermatozoa was significantly correlated with the sperm count and with percentage motility. Sperm count and motility were significantly correlated with each other. The percentages of tapered and amorphous spermatozoa were inversely correlated with the motile sperm count. These results demonstrate that the pattern of spermatozoal morphology observed in men with varicocele is not characteristic or unique to this condition. It appears that abnormality of spermatozoal morphology is commonly associated with diminished sperm count and motility, regardless of the presence or absence of a varicocele.

Humans↗

Semen analysis in testicular cancer and Hodgkin's disease: pre- and post-treatment findings and implications for cryopreservation.

Over a 7-year period, seminal analysis has been performed on 208 patients with testicular tumours, after orchiectomy, but before any other treatment. Only 22% of 54 patients with seminomas, and 29% of 154 patients with teratomas or mixed tumours, had sperm counts exceeding 10 million per ml. Very low sperm counts were observed in some patients who had previously fathered children. Post-treatment sperm counts were done in 117 patients, 80 of whom had received multiple drug chemotherapy: 42 of these men had pre- and post-treatment sperm counts. Overall, 24% of men receiving chemotherapy recovered sperm counts greater than 10 million per ml up to 3 years after therapy. Surprisingly, such recovery was seen in 35% of 23 men with initially poor sperm counts, but in only 26% of 19 with good initial counts. Only 27% of 49 patients with Hodgkin's disease had initial sperm counts of more than 10 million per ml; after chemotherapy only 1 of 29 patients recovered to this level. Only one quarter of these young men had semen which was adequate for cryopreservation. Artificial insemination with semen preserved in liquid nitrogen has been performed in 15 couples: 2 normal babies have been produced and a third pregnancy is progressing normally.

Dysgerminoma↗

Semen quality in Greenland.

OBJECTIVES: To quantify semen quality in the Greenlandic population. STUDY DESIGN: A cross-sectional study including recently proven fertile men from four regions including nine municipalities and one settlement in Greenland. METHODS: The samples were analysed for sperm cell concentrations and motility using standard methods. RESULTS: In total 201 semen samples were collected. The median sperm cell concentration of fertile men in Greenland was 53 x 10(6) sperm cells/ml, with a median sperm cell volume of 3.2 ml, a total sperm count of 185.6 x 10(6) sperm cells and a median motility of 60%. Regionally there were no statistical significant difference in sperm counts, but the sperm cell motility differed among regions, with the lowest sperm cell motility found in semen samples collected on the east coast. Overall sperm cell concentration and total sperm count in Greenland seems to be in the lower range compared to studies from Europe, USA and Japan. CONCLUSION: Within Greenland the sperm cell concentration appears to be similar in different regions, but sperm cell motility tends to be lower in men from the east coast. This may be related to the higher organochlorine exposure level previously demonstrated in this population.

Greenland↗

Comparative analysis of blood plasma epidermal growth factor concentrations, hormonal profiles and semen parameters of fertile and infertile males.

The relationship between male reproductive function and the blood plasma level of epidermal growth factor (EGF) is of interest in the light of the role that circulating EGF appears to play in regulating mouse spermatogenesis. We measured the concentrations of EGF in the blood plasma of 39 fertile men (sperm count > 20 x 10(6)/ml) and compared them with those of 31 infertile men (sperm < 20 x 10(6)/ml). Blood plasma levels of follicle stimulating hormone (FSH), luiteinising hormone (LH), prolactin and testosterone were also determined. The infertile patients had mean blood plasma EGF concentrations of 0.75 +/- 0.10 ug/L. The value was significantly lower than that of the fertile group (1.28 +/- 0.14 ug/L; P < 0.005). There were statistically significant differences between the fertile and infertile groups in sperm count, sperm viability, mean forward progression, testosterone, LH and FSH (P values between 0.0001 and 0.023). There was no significant difference in the prolactin concentrations between the two groups. Although overall average blood plasma EGF concentrations are significantly lower in the infertile males, regression analysis failed to reveal any direct relationships among the various parameters studied.

Adult↗

Follicle-stimulating hormone and human spermatogenesis.

The role of follicle-stimulating hormone (FSH) in the control of spermatogenesis is not well established in any species, including man. We studied the effect of an experimentally-induced, selective FSH deficiency on sperm production in normal men. After a 3-mo control period, five normal men received testosterone enanthate (T) 200 mg i. m. weekly to suppress luteinizing hormone (LH) and FSH, until three successive sperm counts revealed azoospermia or severe oligospermia (sperm counts <3 million/ml). Then, while continuing T, human chorionic gonadotropin (hCG) 5,000 IU i. m. three times weekly was administered simultaneously to replace LH activity, leaving FSH activity suppressed. The effect of the selective FSH deficiency produced by hCG plus T administration on sperm production was determined. Sperm counts (performed twice monthly throughout the study) were markedly suppressed during T administration alone (1.0+/-1.0 million/ml mean+/-SE, compared with 106+/-28 million/ml during the control period, P < 0.001). With the addition of hCG to T, sperm counts returned toward normal (46+/-16 million/ml, P < 0.001 compared with T alone). In two subjects, sperm counts during hCG plus T returned into the individual's control range. Sperm motility and morphology were consistently normal in all men during hCG plus T. Serum FSH levels by RIA were normal (110+/-10 ng/ml) in the control period and were suppressed to undetectable levels (<25 ng/ml) in the T alone and hCG plus T periods. Urinary FSH excretion was markedly suppressed in the T alone (60+/-15 mIU/h-2nd IRP, P < 0.01) and hCG plus T (37+/-9 mIU/h, P < 0.01) periods compared with the control period (334+/-78 mIU/h). We conclude that spermatogenesis as assessed by sperm counts, motilities, and morphologies may be reinitiated and maintained at normal levels in men with undetectable blood FSH levels and urinary excretion of FSH less than that of prepubertal children. This conclusion implies that, although FSH may exert effects on human testicular function, maintenance of normal spermatogenesis and reinitiation of sperm production after short-term suppression by exogenous steroids can occur in spite of nearly absent FSH stimulation.

Adult↗

Seasonal variation and age-related changes in human semen parameters.

Although semen quality has been discussed extensively with regard to age and season in the andrology literature, the results vary and firm conclusions are still outstanding. To investigate seasonal and age-related variations in human semen parameters, we analyzed data that were collected from an andrology clinic population. We performed a retrospective review of 551 semen analysis records collected from 1989 to 2000 from the Vincent Memorial Andrology Laboratory at Massachusetts General Hospital. Semen volume, sperm concentration, total sperm count, motility, total motile sperm, and morphology significantly decreased as age increased. In addition, as age increased, the percentage of sperm with tail defects increased. Sperm concentration was significantly higher in winter (mean 157.9 million/mL) than in fall (mean 119.1 million/mL) (P <.05). The mean percentage of sperm with normal morphology was significantly higher in winter (9.2%) than in summer and spring (7.0% and 7.5%, respectively; P <.05). The mean percentage of sperm with head defects was significantly higher in fall and summer (74.0% and 72.3%, respectively) than in winter (68.6%; P <.05). Seasonal variations were found in sperm concentration and morphology, with higher sperm concentrations in winter than in fall, and a greater percentage of sperm with normal morphology in winter than in spring and summer. Sperm concentration was lowest in the fall, whereas the percentage of sperm with normal morphology was lowest in summer. Semen volume, sperm concentration, total sperm count, motility, total motile sperm, and morphology decreased as age increased.

Adult↗

[Therapeutic evaluation of male infertility--the increase of seminal transferrin level and the improvement in sperm concentration following administration of clomiphene citrate].

This study was designed to evaluate the functional changes of Sertoli cells following the administration of clomiphene citrate for male infertility. Sperm count and seminal transferrin level were measured before and after the treatment in 22 cases of oligozoospermia (sperm count: less than 20 X 10(6)/ml) and 14 cases of subnormal sperm count group (sperm count: 20-30 X 10(6)/ml). Clomiphene citrate was administered per os for more than 3 months consecutively in a dose of 25 mg/day. Seminal transferrin concentration increased more than 1.5 times compared with pre-treatment level in 6 cases (16.7%). Among these patients, sperm count markedly increased (20 X 10(6)/ml or more than the pre-treatment level) in 3 cases (50%) and slightly increased (10 X 10(6)/ml or more than pre-treatment) in 1 case (16.7%). In 30 cases, in which seminal transferrin level did not increase, sperm count markedly increased in 6 cases (20%) and slightly increased in 6 cases (20%). Thus, sperm count was improved more frequently in the cases in which seminal transferrin level remained elevated than the cases with no elevation of serum transferrin level. Serum FSH level of patients whose seminal transferrin level remained elevated after the treatment was significantly higher than that of patients with no elevation of serum transferrin level (mean +/- SD = 32.8 +/- 18.0 mIU/ml v.s. 14.4 +/- 11.7 mIU/ml, respectively). These data suggested that the activation of Sertoli cells may contribute to the increase of sperm count following the administration of clomiphene citrate and the elevated seminal transferrin secretion may be related to the increase of serum FSH level following this treatment.

Clomiphene↗

Adverse testicular effects of some quinolone members in rats.

In the last few years, a marked decrease in male fertility has been reported. Environmental factors were recently suspected for this effect. Among those factors is the misuse of drugs and in particular antibiotics. Quinolones are a group of antibacterial agents with broad-spectrum activity. Testicular impairment of some quinolone members is controversial; a matter which stimulated our attention to investigate the adverse testicular effects of the most familiar quinolone members, namely: ofloxacin, ciprofloxacin and pefloxacin. They were given to rats in doses of 72, 135 and 72 mg kg(-1) day(-1) p.o., respectively, for 15 consecutive days. Ofloxacin was also used to establish a dose-response relationship in doses of 36, 72 and 360 mg kg(-1) day(-1) p.o. for 15 consecutive days. Results revealed that ofloxacin, ciprofloxacin and pefloxacin reduced testicular LDH-X activity by 39.8%, 62.7% and 60.7%, respectively. Moreover, sperm count, motility and daily sperm production were markedly decreased. Ofloxacin induced a dose-dependent decrease in testicular LDH-X activity, sperm count and motility. Furthermore, daily sperm production showed a marked reduction which amounted to 26.1% and 40. 0% following administration of ofloxacin (72, 360 mg kg(-1) day(-1) x 15 days), respectively. Moreover, administration of ofloxacin resulted in marked testicular histopathological changes. It is concluded that, ofloxacin, ciprofloxacin and pefloxacin significantly impaired both testicular function and structure in rats.

Acid Phosphatase↗